title: Rotavirus mRNAS are released by transcript-specific channels in the double-layered viral capsid.
abstract: Rotaviruses are the single most common cause of fatal and severe childhood diarrheal illness worldwide (>125 million cases annually). Rotavirus shares structural and functional features with many viruses, such as the presence of segmented double-stranded RNA genomes selectively and tightly packed with a conserved number of transcription complexes in icosahedral capsids. Nascent transcripts exit the capsid through 12 channels, but it is unknown whether these channels specialize in specific transcripts or simply act as general exit conduits; a detailed description of this process is needed for understanding viral replication and genomic organization. To this end, we developed a single molecule assay for capturing and identifying transcripts extruded from transcriptionally active viral particles. Our findings support a model in which each channel specializes in extruding transcripts of a specific segment that in turn is linked to a single transcription complex. Our approach can be extended to study other viruses and transcription systems.
authors.affiliation: Biological Physics Research Group, Clarendon Laboratory, Department of Physics, University of Oxford, Oxford OX1 3PU, United Kingdom.
authors.forename: Javier
authors.initials: J
authors.lastname: Periz
authors.forename: Cristina
authors.initials: C
authors.lastname: Celma
authors.forename: Bo
authors.initials: B
authors.lastname: Jing
authors.forename: Justin N M
authors.initials: JN
authors.lastname: Pinkney
authors.forename: Polly
authors.initials: P
authors.lastname: Roy
authors.forename: Achillefs N
authors.initials: AN
authors.lastname: Kapanidis
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D002213:Capsid / Q000378:metabolism / Q000502:physiology*; D008253:Macaca mulatta / Q000821:virology*; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009841:Oligonucleotides / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism*; D012401:Rotavirus / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics*
chemical_list: D009841:Oligonucleotides; D012333:RNA, Messenger
keywords: Reoviridae; channel specialization; single molecule hybridization; single-stranded RNA